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Human transcription factor IIIC box B binding subunit
N D L'Etoile1, M L Fahnestock, Y Shen
1Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90024-1570.
Summary
Researchers cloned and sequenced the human TFIIIC alpha subunit, a key protein for RNA polymerase III transcription. This finding is crucial for understanding gene regulation in response to growth factors and viral infections.
Area of Science:
- Molecular Biology
- Gene Regulation
- Biochemistry
Background:
- Transcription factor IIIC (TFIIIC) is essential for RNA polymerase III transcription of tRNA and 5S rRNA genes.
- TFIIIC activity regulation is vital for cellular responses to growth factors and viral infections in mammalian cells.
Purpose of the Study:
- To clone and sequence the full-length cDNA encoding the box B binding subunit of human TFIIIC.
- To functionally characterize the cloned subunit and confirm its role in TFIIIC.
Main Methods:
- Full-length cDNA and genomic DNA sequencing.
- Generation of specific antisera against the encoded protein.
- Electrophoretic mobility shift assays (EMSAs).
- Immunodepletion assays.
Main Results:
- The 243-kDa alpha subunit of human TFIIIC was successfully cloned and sequenced.
- Antisera confirmed the protein's identity as a TFIIIC component by super-shifting DNA-protein complexes and depleting transcriptional activity.
- The human TFIIIC subunit shows limited similarity to its yeast counterpart.
Conclusions:
- The cloned cDNA encodes the functional box B binding subunit of human TFIIIC.
- This research provides a molecular basis for studying TFIIIC regulation in human cells.
- Comparative analysis reveals evolutionary divergence in TFIIIC subunits between humans and yeast.